Method for preparing high-activity pancreatin

A high-activity, pancreatic enzyme technology, applied in the direction of pancreatic enzymes, can solve the problems of large differences in activity and destruction of the three enzymes

Inactive Publication Date: 2013-10-09
SICHUAN UNIV
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  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, the preparation of pancreatin is mostly extracted with low-concentration organic solvents (such as 25% ethanol, 10% acetone, 7.5% isopropanol, etc.), and then precipitated with high-concentration organic solvents. The three enzymes in the pancreatin produced by each process (Trypsin, pancreatic lipase and pancreatic amylase) activities vary greatly, and there is no process with relatively high activities of the three enzymes of trypsin, especially the activity of pancreatic lipase is most likely to be damaged by organic solvents and proteases, so through It is an important measure to optimize the production process of pancreatic enzymes by using mild extraction and precipitation methods and adding suitable protective agents to improve the activity of each enzyme

Method used

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  • Method for preparing high-activity pancreatin
  • Method for preparing high-activity pancreatin
  • Method for preparing high-activity pancreatin

Examples

Experimental program
Comparison scheme
Effect test

example 1

[0020] This example examines the effect of extracting pancreatin with the extract composed of this formula. Add 2% CaCl to 50g pancreatic residue 2 Solution 100ml, stir well, and add 0.5% NH 4 Cl, 0.5% NaCl, 1% sucrose, 1% starch, 1% glycerol and 0.5% Tween 80, while using 4molL -1 Na 2 HPO 4 Adjust pH6.5-7.0. Let cool to 4°C, add 30% PEG 260mL, stir for 30min, place in a refrigerator at 4°C for 1h, siphon the supernatant, centrifuge at 3500rpm for 10min, degrease the precipitate twice with twice the volume of cold acetone, once with ether, and dry to obtain trypsin . The activities of trypsin, pancreatic lipase and pancreatic amylase in pancreatic enzymes were determined by the method of Chinese Pharmacopoeia 2010 edition. with no added NH 4 Cl, sucrose, starch, glycerol and Tween 80 extracts were extracted for comparison. The comparison results are shown in Table 1. The yield of trypsin has increased by 16%, and the activities of the three enzymes have been signific...

example 2

[0023] This example examines NH 4 The stimulant effect of Cl addition on the enzyme activity of pancreatic lipase. Control group except without NH 4 Except for Cl, all the others were operated as described above. results in the removal of NH 4 After Cl, the yield of trypsin, trypsin and amylase hardly changed, and the activity of trypsin decreased by 2.6%. The results are shown in Table 2.

[0024]

example 3

[0026] This example investigates the protective effect of sucrose on pancreatic enzymes. After removing 1% of sucrose from the combined protective agent, the activities of the three enzymes in the obtained pancreatic enzymes are all reduced, trypsin is reduced by 3.7%, pancreatic amylase is reduced by 5.3%, and pancreatic fat The enzyme decreased by 5.6%, and the yield of trypsin did not change significantly (Table 3).

[0027]

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Abstract

The invention provides a method for preparing high-activity pancreatin. The method comprises the following steps of: by adopting porcine pancreas or the rest pancreatic slag generated by extracting insulin through neutral ethanol as a raw material; adding 2% of calcium chloride solution for activating, adding moderate NaCl, NH4Cl, sucrose, starch, glycerinum and tween 80; adjusting pH (Potential of Hydrogen) to 5.5 to 7.0 through 4mol/Lof Na2HPO4; activating for 6 hours at 30 DEG C; dividing size and filtering to remove residues; standing for cooling to reach 4 to 6 DEG C; precipitating through PEG (Polyethylene Glycol)-6000; degreasing through acetone; and drying to obtain the pancreatin. According to the method, the yield of the pancreatin is increased by 16% under the pancreatin extraction environment set in the experiment, and the activity of three enzymes is obviously improved, wherein the activity of trypsin is increased by 13%, the activity of amylopsin is increased by 26%, and the activity of pancreatic lipase is increased by 43%; the activity of the pancreatic lipase, which is easily damaged, is greatly improved.

Description

technical field [0001] The invention relates to a method for preparing high-activity trypsin to ensure that the prepared trypsin has higher yield and activity. Background technique [0002] Animal pancreatin is called "enzyme library" because it contains a variety of enzymes, such as proteolytic enzymes (trypsin, chymotrypsin, etc.), pancreatic lipase, elastase, etc. Pancreatin is a digestive aid drug recorded in Chinese and international pharmacopoeias, used to treat indigestion and digestive disorders caused by liver diseases, etc. [1] . In addition, pancreatin is also used in scientific research as a biological enzyme. At the same time, pancreatin is mostly used in industrial sectors such as leather processing, silk, textiles, printing and dyeing, etc. [2] . [0003] In my country, there were many studies on the preparation of pancreatic enzymes extracted from animal pancreas in the 1980s and 1990s. At present, the preparation of pancreatin is mostly extracted with l...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N9/94
Inventor 崔瑜霞余蓉王欢欢杨继虞
Owner SICHUAN UNIV
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